核磁共振光谱结合选择性约束的多元曲线分辨解析绿豆种子萌发中的代谢变化  

Analysis of Metabolism During Mung Bean Germination by 1H-NMR Combining With MCR-ALS With Selectivity Constraint

作　　者：李响 李中峰[1] 高俊 王霞 张欣[1] 张卓勇[1] LI Xiang;LI Zhong-feng;GAO Jun;WANG Xia;ZHANG Xin;ZHANG Zhuo-yong(Department of Chemistry,Capital Normal University,Beijing 100048,China)

机构地区：[1]首都师范大学化学系,北京100048

出　　处：《光谱学与光谱分析》2020年第10期3010-3014,共5页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金青年科学基金项目(21705112);北京市教育委员会科学研究项目(KM201710028009,KM201910028014);北京市优秀人才培养资助青年骨干个人项目(18540090004);首都师范大学青年创新团队(009175301300)资助。

摘　　要：绿豆作为一种大宗粮食作物,营养丰富,可以直接食用,也可培育为豆芽或制成淀粉及其制品等。研究种子的萌发过程能够为粮食研究提供重要的科学依据和数据基础,为了研究植物在代谢过程中代谢物的变化,分别对绿豆种子在24,36和48 h的萌发样品进行代谢物的提取,通过^1H-NMR测试,分析绿豆在不同萌发时间内代谢物的变化。^1H-NMR除无损、稳定等特点外,尚存在一些不足,例如该方法灵敏度较低,在复杂体系中谱峰会存在严重重叠现象,不易分辨代谢物特征峰,因此需要化学计量学方法对其中的信息进行提取。采用了多元曲线分辨-交替最小二乘法(MCR-ALS)分析NMR的代谢组学数据。MCR为双线性模型,能够将测定的复杂数据矩阵分解并提供各主成分NMR纯光谱和其对应的相对浓度,在代谢组学中能够获得具有实际理化意义的结果,便于检索光谱中对应谱库的代谢物及其浓度变化。仅使用非负约束往往导致双线性模型无法获得唯一解,在本文中,MCR迭代过程除了施加非负约束还使用了选择性约束,对于判断不包含相应代谢物的主成分,可将其^1H-NMR谱图强度约束为零。结果表明,乙酸、胆碱在绿豆萌发前期存在,萌发后期含量呈现升高趋势;蔗糖、缬氨酸、α-酮戊二酸色氨酸、苯丙氨酸、肌苷在萌发之前未见存在,随绿豆萌发含量逐渐增加;O-甲基鲨肌酸、磷酸胆碱、α-葡萄糖在萌发前存在,在萌发初期不断增加,直至萌发后期,因被消耗而含量逐渐降低,表现出先增加后降低的趋势。乳酸、2-氨基-4-氧戊酸、苹果酸、柠檬酸、胆碱、棉籽糖家族寡糖、α-半乳糖、N1-甲基烟酸在绿豆萌发期间一直存在,含量未见明显变化。MCR能够方便地利用先验性质提供NMR中包含的代谢物信息,此方法可推广到不同植物的代谢研究中。The research of the Seed germination progress could provide important scientific information and corresponding database for food crops.As one of the primary agricultural products,mung bean is nutrients-rich and can be eaten directly,manufactured as products or cultured to bean sprouts.In order to study the changes of metabolites in plants during the process of metabolism,we extracted the metabolites from the germination samples of mung bean seeds at 24 h,36 h and 48 h,and analyzed the changes of metabolites in mung bean seeds by 1H-NMR.Except for its non-destructive and stable characteristics,1H-NMR still has some shortcomings of low sensitivity and low resolution in the spectrum.In complex systems,there is overlap in spectrum bands for different compounds,which makes it difficult to give enough information on metabolites.Therefore,chemometric methods are needed to interpret the information in NMR spectroscopy.In this paper,multivariate curve resolution-alternating least squares(MCR-ALS)was used for data processing and analysis of the metabolite data based on NMR.MCR is a bilinear model,which can decompose the given data matrix to the pure spectra and their corresponding relative concentration.MCR can provide physical and chemical meaningful results,which facilitate the analysis of metabolites based on the corresponding spectral library and provide their concentration changes.In this paper,besides the non-negative constraints,selectivity constraint was also used in MCR iteration to reduce the rotation ambiguity in MCR modeling.The changes of metabolites and differential metabolites in mung bean at different germination stages were analyzed.In the determination of samples without certain metabolites,the 1H-NMR spectra are constrained to zero.From the MCR results,we can find that acetate and choline exist in the pre-germination stage of mung bean,and increasing in the late stage of germination.Sucrose,valine,α-ketoglutarate tryptophan,and phenylalanine inosine do not exist in the pre-germination stage,while they g

关 键 词：多元曲线分辨-交替最小二乘 种子萌发 核磁共振氢谱 

分 类 号：O651[理学—分析化学]